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Real Time RT-PCR

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

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quantGenius: implementation of a decision support system for qPCR-based gene quantification.

Špela Baebler¹, Miha Svalina², Marko Petek²

¹Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, 1000, SI, Slovenia. spela.baebler@nib.si.

BMC Bioinformatics

|May 27, 2017

Summary

Scientists can now achieve reliable nucleic acid quantification with quantGenius, a new web application. This tool offers robust quality control and decision support for accurate quantitative PCR (qPCR) data analysis.

Keywords:

Decision support system Nucleic acid quantification Quantitative PCR Quantitative molecular biology Web application

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Area of Science:

Molecular Biology
Bioinformatics

Background:

Quantitative molecular biology presents challenges due to inconsistent error control in results.
Quantitative analysis and interpretation of quantitative PCR (qPCR) data are complex.
High-throughput qPCR platforms necessitate tools for robust, reliable, and fast nucleic acid quantification.

Purpose of the Study:

To develop an open-access web application for reliable qPCR-based nucleic acid quantification.
To provide a tool that integrates quality control (QC) decision support for qPCR data analysis.

Main Methods:

Developed "quantGenius", an open-access web application for qPCR data analysis.
Implemented a user-guided QC-based decision support system considering pipetting errors, assay efficiencies, and PCR inhibition.
Quantification uses the standard curve approach with reference gene normalization.

Main Results:

quantGenius offers machine- and chemistry-independent data import and analysis.
The application provides user-guided QC decision support for reliable qPCR results.
Intermediate calculations and final results are exportable for further analysis and visualization.

Conclusions:

quantGenius is a unique qPCR data analysis tool integrating QC-based decision support.
The application aids scientists in obtaining reliable results for biologically meaningful data interpretation.